Bakshi Arkadipta, Gilroy Simon
Department of Botany, University of Wisconsin-Madison, Madison, WI, United States.
Department of Botany, University of Wisconsin-Madison, Madison, WI, United States.
Methods Enzymol. 2023;680:461-491. doi: 10.1016/bs.mie.2022.08.043. Epub 2022 Nov 11.
Flooding represents an environmental stress that has widespread effects on plants in natural ecosystems as well as causing major crop losses. As climate change leads to more severe weather extremes, severe flooding events are likely to become even more frequent. Thus, there is intense interest in understanding how flooding affects plants and in identifying cellular and molecular targets for engineering flood resilience into crop species. Such research requires well controlled, highly reproducible flooding protocols for use in the laboratory. However, there are many ways that a plant can be flooded. For example, waterlogging of the soil, where water levels reach the soil surface, generally generates a hypoxic environment around the root system. In contrast, full submergence of the plant adds effects on the aerial organs such as impaired photosynthesis from the combination of lowered CO availability and the reduced light penetration into often turbid flood waters. In this chapter, approaches to imposing controlled flooding conditions to the model plant Arabidopsis thaliana are discussed. A series of straight-forward assays are then described to document the effects of stress-related changes in growth patterns, pigment accumulation and levels of oxidative stress. These assays are complemented by monitoring the expression of a series of molecular markers of flood response by qPCR.
洪水是一种环境胁迫,它不仅会对自然生态系统中的植物产生广泛影响,还会导致农作物严重减产。随着气候变化导致极端天气愈发严重,严重的洪水事件可能会变得更加频繁。因此,人们对了解洪水如何影响植物以及确定将洪水恢复力工程化到作物品种中的细胞和分子靶点有着浓厚的兴趣。此类研究需要在实验室中使用控制良好、高度可重复的洪水试验方案。然而,使植物遭受水淹的方式有很多种。例如,当水位达到土壤表面时,土壤渍水通常会在根系周围产生缺氧环境。相比之下,植物完全被淹没会对地上器官产生额外影响,比如由于二氧化碳供应降低以及光线穿透常常浑浊的洪水而减少,导致光合作用受损。在本章中,将讨论对模式植物拟南芥施加可控水淹条件的方法。随后将描述一系列简单的试验,以记录与胁迫相关的生长模式变化、色素积累和氧化应激水平的影响。通过定量聚合酶链反应(qPCR)监测一系列洪水响应分子标记的表达,对这些试验起到补充作用。